title: Purification to apparent homogeneity of murine interleukin 1.
abstract: Interleukin 1 was purified to apparent homogeneity from the culture fluid of P388D1 cell line macrophages by a procedure involving ammonium sulfate fractionation, phenyl Sepharose and Ultrogel AcA54 chromatographies, and preparative isoelectrofocusing. The interleukin 1 was purified more than 300-fold to a specific activity of approximately 1 X 10(6) U/mg protein. The purified interleukin 1 exhibited microheterogeneity. Three charge species, which we have termed alpha, beta, gamma, were resolved by Tris-glycinate discontinuous polyacrylamide gel electrophoresis. The partial amino acid content of the alpha species of interleukin 1 was determined. On the basis of the purification data, it appears that interleukin 1 is biologically active within the 10(-11) to 10(-10) M concentration range. The success of the purification was due in part to the use of a superinduction protocol that resulted in the production of interleukin 1 at a rate of approximately 1 microgram/24 hr/4 X 10(6) P388D1 cells.
mesh_terms: D000596:Amino Acids; D000818:Animals; D002087:Butyrates / Q000494:pharmacology; D002417:Cattle; D002460:Cell Line; D005591:Chemical Fractionation; D002847:Chromatography, Agarose; D002850:Chromatography, Gel; D003513:Cycloheximide / Q000494:pharmacology; D003609:Dactinomycin / Q000494:pharmacology; D004591:Electrophoresis, Polyacrylamide Gel; D005260:Female; D007375:Interleukin-1; D007525:Isoelectric Focusing; D008264:Macrophages; D051379:Mice; D011247:Pregnancy; D011506:Proteins / Q000302:isolation & purification*; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology
chemical_list: D000596:Amino Acids; D002087:Butyrates; D007375:Interleukin-1; D011506:Proteins; D003609:Dactinomycin; D003513:Cycloheximide; D013755:Tetradecanoylphorbol Acetate